Checkpoint Kinase 1 Stimulates Endogenous Cardiomyocyte Renewal and Cardiac Repair by Binding to Pyruvate Kinase Isoform M2 C-Domain and Activating Cardiac Metabolic Reprogramming in a Porcine Model of Myocardial Ischemia/Reperfusion Injury.

Autor: Wei TW; Department of Cardiology The First Affiliated Hospital of Nanjing Medical University Nanjing China., Shan TK; Department of Cardiology The First Affiliated Hospital of Nanjing Medical University Nanjing China., Wang H; Department of Cardiology The First Affiliated Hospital of Nanjing Medical University Nanjing China., Chen JW; Department of Cardiology The First Affiliated Hospital of Nanjing Medical University Nanjing China., Yang TT; Department of Cardiology The First Affiliated Hospital of Nanjing Medical University Nanjing China., Zhou LH; Department of Cardiology The First Affiliated Hospital of Nanjing Medical University Nanjing China., Zhao D; Department of Cardiology The First Affiliated Hospital of Nanjing Medical University Nanjing China., Sun JT; Department of Cardiology The First Affiliated Hospital of Nanjing Medical University Nanjing China., Wang SB; Department of Cardiology The First Affiliated Hospital of Nanjing Medical University Nanjing China., Gu LF; Department of Cardiology The First Affiliated Hospital of Nanjing Medical University Nanjing China., Du C; Department of Cardiology The First Affiliated Hospital of Nanjing Medical University Nanjing China., Jiang QQ; Department of Cardiology The First Affiliated Hospital of Nanjing Medical University Nanjing China., Sun R; Department of Cardiology The First Affiliated Hospital of Nanjing Medical University Nanjing China., Wang QM; Department of Cardiology The First Affiliated Hospital of Nanjing Medical University Nanjing China., Kong XQ; Department of Cardiology The First Affiliated Hospital of Nanjing Medical University Nanjing China., Lu XH; Department of Cardiovascular Surgery The First Affiliated Hospital of Nanjing Medical University Nanjing China., Sun HL; Department of Cardiovascular Surgery The First Affiliated Hospital of Nanjing Medical University Nanjing China., Xu Y; Department of Radiology The First Affiliated Hospital of Nanjing Medical University Nanjing China., Xie LP; Key Laboratory of Cardiovascular and Cerebrovascular Medicine Key Laboratory of Targeted Intervention of Cardiovascular Disease Collaborative Innovation Center for Cardiovascular Disease Translational Medicine Nanjing Medical University Nanjing China., Gu AH; State Key Laboratory of Reproductive Medicine School of Public Health Nanjing Medical University Nanjing China., Chen F; Department of Biostatistics School of Public Health China International Cooperation Center for Environment and Human Health Nanjing Medical University Nanjing China., Ji Y; Key Laboratory of Cardiovascular and Cerebrovascular Medicine Key Laboratory of Targeted Intervention of Cardiovascular Disease Collaborative Innovation Center for Cardiovascular Disease Translational Medicine Nanjing Medical University Nanjing China., Guo XJ; State Key Laboratory of Reproductive Medicine Department of Histology and Embryology Nanjing Medical University Nanjing China., Wang LS; Department of Cardiology The First Affiliated Hospital of Nanjing Medical University Nanjing China.
Jazyk: angličtina
Zdroj: Journal of the American Heart Association [J Am Heart Assoc] 2024 Jul 02; Vol. 13 (13), pp. e034805. Date of Electronic Publication: 2024 Jun 27.
DOI: 10.1161/JAHA.124.034805
Abstrakt: Background: The regenerative capacity of the adult mammalian hearts is limited. Numerous studies have explored mechanisms of adult cardiomyocyte cell-cycle withdrawal. This translational study evaluated the effects and underlying mechanism of rhCHK1 (recombinant human checkpoint kinase 1) on the survival and proliferation of cardiomyocyte and myocardial repair after ischemia/reperfusion injury in swine.
Methods and Results: Intramyocardial injection of rhCHK1 protein (1 mg/kg) encapsulated in hydrogel stimulated cardiomyocyte proliferation and reduced cardiac inflammation response at 3 days after ischemia/reperfusion injury, improved cardiac function and attenuated ventricular remodeling, and reduced the infarct area at 28 days after ischemia/reperfusion injury. Mechanistically, multiomics sequencing analysis demonstrated enrichment of glycolysis and mTOR (mammalian target of rapamycin) pathways after rhCHK1 treatment. Co-Immunoprecipitation (Co-IP) experiments and protein docking prediction showed that CHK1 (checkpoint kinase 1) directly bound to and activated the Serine 37 (S37) and Tyrosine 105 (Y105) sites of PKM2 (pyruvate kinase isoform M2) to promote metabolic reprogramming. We further constructed plasmids that knocked out different CHK1 and PKM2 amino acid domains and transfected them into Human Embryonic Kidney 293T (HEK293T) cells for CO-IP experiments. Results showed that the 1-265 domain of CHK1 directly binds to the 157-400 amino acids of PKM2. Furthermore, hiPSC-CM (human iPS cell-derived cardiomyocyte) in vitro and in vivo experiments both demonstrated that CHK1 stimulated cardiomyocytes renewal and cardiac repair by activating PKM2 C-domain-mediated cardiac metabolic reprogramming.
Conclusions: This study demonstrates that the 1-265 amino acid domain of CHK1 binds to the 157-400 domain of PKM2 and activates PKM2-mediated metabolic reprogramming to promote cardiomyocyte proliferation and myocardial repair after ischemia/reperfusion injury in adult pigs.
Databáze: MEDLINE
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